Methods and compositions for the treatment of pre-diabetes, diabetes and metabolic syndrome

ABSTRACT

The disclosure provides thiol-containing alkyl fatty acid and vitamin D compound formulations for intra-venous, parenteral or oral administration. The compositions of the present technology have optimal controlled bioavailability and are useful for treating metabolic dysfunctions such as pre-diabetes, Metabolic Syndrome and diabetes. Also provided are methods of treatment comprising the daily administration of the disclosed thiol-containing alkyl fatty acid formulations alone or in combination with the disclosed vitamin D compound formulations.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of U.S. application Ser. No.15/521,216 filed Apr. 21, 2017, which is a U.S. National PhaseApplication of International Application No. PCT/US2015/057382 filedOct. 26, 2015, which claims the benefit of and priority to U.S.Application No. 62/068,028, filed Oct. 24, 2014, each of which isincorporated herein by reference in its entirety.

TECHNICAL FIELD

The present disclosure relates generally to compositions comprising acombination of synthetic chemical agents and herbal extracts asnutritional and dietary supplements and their methods of use for thetreatment of metabolic dysfunction.

BACKGROUND

Much attention has been paid to the concept of “we are what we eat.”While at a superficial level this would be reasonable, mammals are infact much more dependent on what they can (i) digest in the gut, (ii)absorb into the blood stream, (iii) deliver to organs, tissues, cellsand organelles in a form that it useful in meeting metabolic needs. Bothcatabolic and anabolic processes are required to support healthyfunction. The transport of nutrients into cells and their distributioninto the appropriate organelles for metabolism is highly regulated butyet is responsive to dietary changes. Mitochondria play a central rolein the production of ATP, the formation of biosynthetic intermediates aswell as the production of signal transduction molecules involved ingrowth and differentiation and homeostasis.

Changes in these processes as a result of genetics, exposure topathogens or dietary extremes can stress the body to the point ofdisease manifestation. Excessive consumption of processed foods, sugarand inappropriate types and levels of fats may contribute to a varietyof ailments including, but not limited to, diabetes, cardiovasculardisease, gout, and inflammatory disease.

Nutritional supplements can play an important role in reversing orpreventing these diseases. In many instances, the bioavailability of onenutrient is required along with others for the optimal treatment andmaintenance of health. In an age of highly processed foods, keynutrients such as vitamins and essential enzyme cofactors, may belacking or may be destroyed during preparation. However, merereplacement of these missing nutrients by consuming tablets, powders orcapsules containing one or more of these nutrients is insufficientbecause metabolism is a concert of activities and each nutrient,cofactor and enzyme has specific temporal, spatial and structuralrequirements in the context of these orchestrated metabolic events. Itis thus not only a question of what we eat, digest and can absorb in ausable form, i.e. the collective “what” but also “when and where.”

Insulin receptors are desensitized and the ability to clear glucose fromthe blood is compromised in patients with Metabolic Syndrome,pre-diabetes, or diabetes. Given the severe health risks associated withpre-diabetes, diabetes and Metabolic Syndrome, there is a need for safeand effective therapeutics.

SUMMARY

The present disclosure provides alkyl fatty acid formulations andvitamin D compound formulations, selected on the basis ofnon-anticipated synergies and wherein at least two, at least three, orat least four or more of the individual components of thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations of the present technology have overlapping oralbioavailability. The formulations of the present technology permit cellsto have a contemporaneous degree of exposure to the components of theformulations disclosed herein that is otherwise cost prohibitive orimpractical by the dosing of individual components independently. Forexample, depending on the dosage and the form of the medicament (e.g., acapsule, tablet, or powder) the requisite numbers of units of eachcomponent to be ingested may be difficult. Each preparation of anindividual ingredient will have its own dissolution kinetics, thusaffecting the level of bioavailability and plasma pharmacokinetics. Anadditional concern is that the levels of fillers or excipientsmultiplied by the numbers of capsules or tablets can contribute totoxicities within patients.

In one aspect, the present disclosure provides a thiol-containing alkylfatty acid formulation comprising a thiol-containing alkyl fatty acid, aN-acetyl-L-carnitine derivative, an antioxidant, and at least one aminoacid, wherein the thiol-containing alkyl fatty acid is lipoic acid oralpha-lipoic acid.

In some embodiments, the thiol-containing alkyl fatty acid is a mixtureof Rand S isomers of alpha-lipoic acid. Additionally or alternatively,in some embodiments of the thiol-containing alkyl fatty acidformulation, the N-acetyl-L-carnitine derivative is L-carnitine.Additionally or alternatively, in certain embodiments of thethiol-containing alkyl fatty acid formulation, the antioxidant isselected from the group consisting of CoQ10, quercetin, and otherplant-derived flavonoids. Additionally or alternatively, in someembodiments of the thiol-containing alkyl fatty acid formulation, the atleast one amino acid is one or more of the amino acids selected from thegroup consisting of propionyl L-carnitine, L-tartarate, L-glutamine,L-theanine and L-arginine.

In some embodiments of the thiol-containing alkyl fatty acidformulation, the thiol-containing alkyl fatty acid is alpha-lipoic acid,the N-acetyl-L-carnitine derivative is L-carnitine, the antioxidant isCoQ10 and the amino acids are L-glutamine and L-arginine. In someembodiments, the ratio of alpha-lipoic acid to L-carnitine is 1.5:1.Additionally or alternatively, in some embodiments, the ratio ofalpha-lipoic acid to CoQ10 is 6:1. Additionally or alternatively, incertain embodiments, the ratio of alpha-lipoic acid to L-glutamine is1:1.25. Additionally or alternatively, in some embodiments, the ratio ofalpha-lipoic acid to L-arginine is 1:2.5.

In any of the above embodiments, the thiol-containing alkyl fatty acidformulation further comprises one or more of melatonin, proteins,peptides, protease inhibitors, trace elements, botanical extracts, andcarbohydrates. In some embodiments of the thiol-containing alkyl fattyacid formulation, the protein is serratiopeptidase. Additionally oralternatively, in some embodiments of the thiol-containing alkyl fattyacid formulation, the trace element is indium sulfate. Additionally oralternatively, in some embodiments of the thiol-containing alkyl fattyacid formulation, the protease inhibitor is soybean trypsin inhibitor,leupeptin, or pepstatin. Additionally or alternatively, in someembodiments of the thiol-containing alkyl fatty acid formulation, thebotanical extracts are one or more plant extracts selected from thegroup consisting of eggplant extract, resveratrol-containing extractssuch as grape and grape seed extracts, guarana extract, purified orsynthetic whole-plant cannabis extract, vanilla extract, cinnamon,nutmeg, cloves, and turmeric. Additionally or alternatively, in someembodiments of the thiol-containing alkyl fatty acid formulation, thecarbohydrate is selected from the group consisting of beta-glucans,fiber extracts, fruit pulps, simple and complex sugars. In someembodiments, the beta-glucan is -1,3-glucan or -1,6-glucan. Additionallyor alternatively, in some embodiments of the thiol-containing alkylfatty acid formulation, the formulation comprises 600 mg alpha-lipoicacid, 400 mg L-carnitine, 100 mg CoQ10, 750 mg L-glutamine and 1500 mgL-arginine.

In another aspect, the present disclosure provides a vitamin D compoundformulation comprising a vitamin D compound, a sulfur donor and one ormore sedatives or sleep promoting agents, wherein the vitamin D compoundis cholecalciferol (D3) or ergocalciferol (D2). In some embodiments ofthe vitamin D compound formulation, the sulfur donor ismethylsulfonylmethane. Additionally or alternatively, in someembodiments of the vitamin D compound formulation, the one or moresedatives or sleep promoting agents are selected from the groupconsisting of melatonin, L-theanine, chamomile extract, valerianextract, 5-hydroxytryptamine (5-HT), SHTP (L-5-hydroxytryptophan),D-phenylalanine, L-phenylalanine, or a mixture of D- andL-phenylalanine.

In any of the above embodiments, the vitamin D compound formulationfurther comprises melatonin, proteins, peptides, protease inhibitors,trace elements, botanical extracts, and carbohydrates. In someembodiments of the vitamin D compound formulation, the protein isserratiopeptidase. Additionally or alternatively, in some embodiments ofthe vitamin D compound formulation, the trace element is indium sulfate.Additionally or alternatively, in some embodiments of the vitamin Dcompound formulation, the protease inhibitor is soybean trypsininhibitor, leupeptin, or pepstatin. Additionally or alternatively, insome embodiments of the vitamin D compound formulation, the botanicalextracts are one or more plant extracts selected from the groupconsisting of eggplant extract, resveratrol-containing extracts such asgrape and grape seed extracts, guarana extract, purified or syntheticwhole-plant cannabis extract, vanilla extract, cinnamon, nutmeg, cloves,and turmeric. Additionally or alternatively, in some embodiments of thevitamin D compound formulation, the carbohydrate is selected from thegroup consisting of beta-glucans, fiber extracts, pulps, simple andcomplex sugars. In some embodiments, the beta-glucan is -1,3-glucan or-1,6-glucan. Additionally or alternatively, in some embodiments of thevitamin D compound formulation, the formulation comprises 1000 IUvitamin D, 400 mg L-theanine, 400 mg methylsulfonylmethane, 50 mg 5-HTP,3 mg melatonin, 600 mg valerian root powder, and 500 mg chamomilepowder.

Additionally or alternatively, any of the formulations disclosed hereinmay be provided as an oral solid medicament in the form of tablets,capsules, nanoparticles, or nano-emulsions for immediate or sustainedrelease. The formulations of the present technology may comprise aplurality of crystalline structures. The formulations can also containadditional ingredients or encapsulations that establish the releaseprofile of each or the collective ingredient(s) for instantaneousrelease, or sustained release. In a further embodiment, the oral soliddosage form can be enterically coated so as to prevent gastricirritation in the subject. Alternatively, the formulations of thepresent technology can be either fully dissolved or prepared as anemulsion.

In one aspect, the present disclosure provides methods for treating orpreventing pre-diabetes, diabetes or Metabolic Syndrome, and/or treatingor preventing the signs or symptoms of pre-diabetes, diabetes orMetabolic Syndrome in a subject in need thereof comprising administeringto the subject a therapeutically effective amount of at least one of theformulations disclosed herein, thereby resulting in the prevention ortreatment of one or more signs or symptoms of pre-diabetes, diabetes orMetabolic Syndrome. In some embodiments of the method, the subjectexhibits a reduction in Body Mass Index (BMI) after administration of atleast one of the formulations disclosed herein. In some embodiments ofthe method, the subject exhibits a reduction in glycated hemoglobinlevels (HbA1c) after administration of at least one of the formulationsdisclosed herein. In some embodiments of the method, the weight statusof the subject shifts from overweight to normal weight afteradministration of at least one of the formulations disclosed herein.

In some embodiments of the method, the signs or symptoms ofpre-diabetes, diabetes or Metabolic Syndrome include one or more of bodyweight, fasting glucose/insulin/free fatty acid, glucose tolerance(OGTT), muscle insulin sensitivity, markers of insulin signaling (e.g.,Akt-P, IRS-P), serum triglyceride levels, HDL and LDL cholesterollevels, blood pressure, serum fibrinogen or plasminogen activatorinhibitor levels, levels of C-reactive protein, insulin resistance,glucose intolerance, elevated levels of glycosylated blood proteins(e.g., hemoglobin).

In another aspect, the present disclosure provides methods for treatinga disease or condition characterized by elevated glycated hemoglobinlevels (HbA1c) in a subject in need thereof comprising administering tothe subject a therapeutically effective amount of at least one of theformulations disclosed herein. In some embodiments of the method, thedisease or condition characterized by elevated glycated hemoglobinlevels (HbA1c) is selected from the group consisting of pre-diabetes,diabetes, Metabolic Syndrome, insulin resistance, glucose intolerance orcardiovascular disease, and stroke. In some embodiments, the fastingplasma glucose level (FPG) of the subject prior to administration of atleast one of the formulations disclosed herein is at least 5.6 mmol/L.In some embodiments of the method, the subject exhibits a reduction inglycated hemoglobin levels (HbA1c) after administration of at least oneof the formulations disclosed herein.

In some embodiments of the methods of the present technology, thesubject is human.

Additionally or alternatively, in some embodiments of the methods of thepresent technology, the formulations of the present technology areadministered orally, topically, intranasally, systemically,intravenously, subcutaneously, intraperitoneally, intradermally,intraocularly, iontophoretically, transmucosally, or intramuscularly.

In some embodiments of the methods of the present technology, at leastone of the formulations of the present technology is administered dailyfor 1 week or more. In some embodiments of the methods of the presenttechnology, at least one of the formulations of the present technologyis administered daily for 2 weeks or more. In some embodiments of themethods of the present technology, at least one of the formulations ofthe present technology is administered daily for 3 weeks or more. Insome embodiments of the methods of the present technology, at least oneof the formulations of the present technology is administered daily for4 weeks or more. In some embodiments of the methods of the presenttechnology, at least one of the formulations of the present technologyis administered daily for 6 weeks or more. In some embodiments of themethods of the present technology, at least one of the formulations ofthe present technology is administered daily for 12 weeks or more.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows the standard curve and correlation between HbA1c levels(%) and estimated average blood glucose (mg/dL). The relationshipbetween HbA1c and estimated average blood glucose (eAG) is described bythe formula 28.7×HbA1c-46.7=eAG. FIG. 1B depicts the correlation betweenHbA1c levels (%) and estimated average blood glucose (mg/dL).

FIG. 1B is adapted from the American Diabetes Association website and isavailable at professional.diabetes.org/glucosecalculator.aspx.

DETAILED DESCRIPTION

It is to be appreciated that certain aspects, modes, embodiments,variations and features of the present technology are described below invarious levels of detail in order to provide a substantial understandingof the present technology. The definitions of certain terms as used inthis specification are provided below. Unless defined otherwise, alltechnical and scientific terms used herein generally have the samemeaning as commonly understood by one of ordinary skill in the art towhich this present technology belongs.

As used in this specification and the appended claims, the singularforms “a”, “an” and “the” include plural referents unless the contentclearly dictates otherwise. For example, reference to “a cell” includesa combination of two or more cells, and the like.

All numerical designations, e.g., pH, temperature, time, concentrationand molecular weight, including ranges, are approximations which arevaried (+) or (−) by increments of 1.0 or 0.1, as appropriate, oralternatively by a variation of +/−10%, or alternatively 5% oralternatively 2%. It is to be understood, although not always explicitlystated, that all numerical designations are preceded by the term“about”.

As used herein, the “administration” of an agent, drug, or formulationto a subject includes any route of introducing or delivering to asubject a compound to perform its intended function. Administration canbe carried out by any suitable route, including orally, intranasally,parenterally (intravenously, intramuscularly, intraperitoneally, orsubcutaneously), systemically, intradermally, intraocularly,iontophoretically, transmucosally, intramuscularly, or topically.Administration includes self-administration and the administration byanother.

As used herein, the term “amino acid” includes naturally-occurring aminoacids and synthetic amino acids, as well as amino acid analogs and aminoacid mimetics that function in a manner similar to thenaturally-occurring amino acids. Naturally-occurring amino acids arethose encoded by the genetic code, as well as those amino acids that arelater modified, e.g., hydroxyproline, y-carboxyglutamate, andO-phosphoserine. Amino acid analogs refers to compounds that have thesame basic chemical structure as a naturally-occurring amino acid, i.e.,an a-carbon that is bound to a hydrogen, a carboxyl group, an aminogroup, and an R group, e.g., homoserine, norleucine, methioninesulfoxide, methionine methyl sulfonium. Such analogs have modified Rgroups (e.g., norleucine) or modified peptide backbones, but retain thesame basic chemical structure as a naturally-occurring amino acid. Aminoacid mimetics refers to chemical compounds that have a structure that isdifferent from the general chemical structure of an amino acid, but thatfunctions in a manner similar to a naturally-occurring amino acid. Aminoacids can be referred to herein by either their commonly known threeletter symbols or by the one-letter symbols recommended by the IUPAC-IUBBiochemical Nomenclature Commission.

As used herein, the term “effective amount” refers to a quantitysufficient to achieve a desired therapeutic and/or prophylactic effect,e.g., an amount which results in the reduction of insulin resistance,reduction of glucose intolerance and reduction of levels of glycosylatedblood proteins such as hemoglobin, or which results in partial or fullamelioration of one or more symptoms of diabetes, pre-diabetes andMetabolic Syndrome. In the context of therapeutic or prophylacticapplications, in some embodiments, the amount of a formulationadministered to the subject will depend on the type, degree, andseverity of the disease and on the characteristics of the individual,such as general health, age, sex, body weight and tolerance to drugs.The skilled artisan will be able to determine appropriate dosagesdepending on these and other factors. In the methods described herein,the compositions of the present technology may be administered to asubject having one or more signs, symptoms, or risk factors of diabetes,pre-diabetes and Metabolic Syndrome, including, but not limited to,e.g., body weight, fasting glucose/insulin/free fatty acid, glucosetolerance (OGTT), muscle insulin sensitivity, markers of insulinsignaling (e.g., Akt-P, IRS-P), serum triglyceride levels, HDL and LDLcholesterol levels, blood pressure, serum fibrinogen or plasminogenactivator inhibitor levels, levels of C-reactive protein, insulinresistance, glucose intolerance, elevated levels of glycosylated bloodproteins (e.g., hemoglobin) etc. For example, a “therapeuticallyeffective amount” of the formulations includes levels at which thepresence, frequency, or severity of one or more signs, symptoms, or riskfactors of diabetes, pre-diabetes and Metabolic Syndrome are reduced oreliminated. In some embodiments, a therapeutically effective amountreduces or ameliorates the physiological effects of diabetes,pre-diabetes and Metabolic Syndrome, and/or the risk factors ofdiabetes, pre-diabetes and Metabolic Syndrome, and/or the likelihood ofdeveloping diabetes, pre-diabetes and Metabolic Syndrome.

As used herein, the terms “polypeptide,” “peptide,” and “protein” areused interchangeably herein to mean a polymer comprising two or moreamino acids joined to each other by peptide bonds. Polypeptide refers toboth short chains, commonly referred to as peptides, glycopeptides oroligomers, and to longer chains, generally referred to as proteins.Polypeptides may contain amino acids other than the 20 gene-encodedamino acids. Polypeptides include amino acid sequences modified eitherby natural processes, such as post-translational processing, or bychemical modification techniques that are well known in the art.

As used herein, the terms “subject,” “individual,” or “patient” can bean individual organism, a vertebrate, a mammal, or a human.

A “synergistic therapeutic effect” refers to a greater-than-additivetherapeutic effect which is produced by a combination of at least twotherapeutic agents, and which exceeds that which would otherwise resultfrom the individual administration of the agents. For example, lowerdoses of one or more therapeutic agents may be used in treatingpre-diabetes, diabetes or Metabolic Syndrome, resulting in increasedtherapeutic efficacy and decreased side-effects.

“Treating” or “treatment” as used herein covers the treatment of adisease or disorder described herein, in a subject, such as a human, andincludes: (i) inhibiting a disease or disorder, i.e., arresting itsdevelopment; (ii) relieving a disease or disorder, i.e., causingregression of the disorder; (iii) slowing progression of the disorder;and/or (iv) inhibiting, relieving, or slowing progression of one or moresymptoms of the disease or disorder.

As used herein, “prevention” or “preventing” of a disorder or conditionrefers to a compound that, in a statistical sample, reduces theoccurrence of the disorder or condition in the treated sample relativeto an untreated control sample, or delays the onset of one or moresymptoms of the disorder or condition relative to the untreated controlsample. As used herein, preventing pre-diabetes, diabetes or MetabolicSyndrome includes preventing or delaying the initiation of pre-diabetes,diabetes or Metabolic Syndrome. As used herein, prevention ofpre-diabetes, diabetes or Metabolic Syndrome also includes preventing arecurrence of one or more signs or symptoms of pre-diabetes, diabetes orMetabolic Syndrome.

It is also to be appreciated that the various modes of treatment orprevention of medical conditions as described herein are intended tomean “substantial,” which includes total but also less than totaltreatment or prevention, and wherein some biologically or medicallyrelevant result is achieved.

All metabolic processes either produce or require energy. In mammals,“energy” is derived from glucose, fats and other fuel sources and isconverted to and transported as ATP. Energy production involves acoordination of cellular events at the macro and micro levels. Thecellular machinery necessary for processing fuel into energy andsynthetic intermediates must be present and in good working order. Fuelsmust be transported to where they need to go and in a form suitable forprocessing. The right substrates must contact the proper cellularmachinery at the right time and place, and the movement of substratesand byproducts in and out of cells and organs must be regulated.

All biological activities are dependent on the availability of ATP.While ATP can be produced in the cell cytosol, the primary point ofproduction is within the mitochondria. The absence of nutrients requiredfor mitochondrial function, or the presence of cellular deformation orderegulation can lead to pathology.

It is desirable to have medicaments in a novel combination with lipoicacid wherein the combination demonstrates synergistic effects in bothmechanism of action and clinical efficacy.

In some embodiments, the present disclosure provides thiol-containingalkyl fatty acid formulations that improve glucose uptake and theoxidation of nutrients, either alone or in combination with a vitamin Dcompound formulation. In some embodiments, the thiol-containing alkylfatty acid formulations of the present technology, alone or incombination with the vitamin D compound formulations of the presenttechnology are useful in treating or preventing insulin resistance in asubject (including, but not limited to, a subject diagnosed withpre-diabetes, diabetes or Metabolic Syndrome).

The present disclosure provides compositions and methods for treatingpre-diabetes, diabetes, Metabolic Syndrome, insulin resistance, glucoseintolerance and elevated glycated hemoglobin levels (HbA1c) in a subjectin need thereof. The formulations of the present technology permit cellsto have a contemporaneous degree of exposure to the components of theformulations disclosed herein that is otherwise cost prohibitive orimpractical by the dosing of individual components independently. Forexample, depending on the dosage and the form of the medicament (e.g., acapsule, tablet, or powder) the requisite numbers of units of eachcomponent to be ingested may be difficult. Each preparation of anindividual ingredient will have its own dissolution kinetics, thusaffecting the level of bioavailability and plasma pharmacokinetics. Anadditional concern is that the levels of fillers or excipientsmultiplied by the numbers of capsules or tablets can contribute totoxicities within patients.

Pre-Diabetes, Diabetes and Metabolic Syndrome

The diabetic, Metabolic Syndrome and the pre-diabetic states have oftenbeen characterized with respect to insulin, e.g., an insufficient amountof insulin, ineffective insulin, insulin resistance and insulindesensitization. Normal glucose tolerance in healthy control subjects isdefined as a Fasting Plasma Glucose level (FPG) of <5.6 mmol/L and a 2hour plasma glucose level (PG) of <7.8 mmol/L in response to a 75 g oralglucose tolerance test (OGTT). Nathan et al., Impaired Fasting Glucoseand Impaired Glucose Tolerance: Implications for Care. A ConsensusStatement from the American Diabetes Association. Diabetes Care 30:753-9(2007). Diabetes is defined as a FPG 2≥7.0 mmol/L or a 2 hour PG 2≥11.1mmol/L during an OGTT. Pre-diabetes is an intermediate state of alteredglucose metabolism with a heightened risk of developing type 2 diabetesand other associated complications. There are two categories ofpre-diabetes: (a) impaired fasting glucose (IFG), defined by a FPG 2≥5.6mmol/L but <7.0 mmol/L and (b) impaired glucose tolerance (IGT), definedby a FPG <7.0 mmol/L with a 2 h PG during an OGTT 2≥7.8 mmol/L and <11.1mmol/L. IFG and IGT can occur as mutually exclusive conditions (isolatedIFG or isolated IGT) or they can occur in combination (combined IFG andIGT). Nathan et al., Impaired Fasting Glucose and Impaired GlucoseTolerance: Implications for Care. A Consensus Statement from theAmerican Diabetes Association. Diabetes Care 30:753-9 (2007).

Metabolic Syndrome, also known as syndrome X and dysmetabolic syndrome,is a collection of health disorders or risks that increase the chance ofdeveloping heart disease, stroke, and diabetes. Metabolic Syndrome mayinclude a variety of underlying metabolic phenotypes, including insulinresistance and/or obesity predisposition phenotypes. Metabolic Syndromeis often characterized by any of a number of metabolic disorders or riskfactors, which are generally considered to most typify MetabolicSyndrome when more than one of these factors are present in a singleindividual. The factors include: central obesity (disproportionate fattissue in and around the abdomen), atherogenic dyslipidemia (whichincludes a family of blood fat disorders including, e.g., hightriglycerides, low HDL cholesterol, and high LDL cholesterol that canfoster plaque buildups in the vascular system, including artery walls),high blood pressure, insulin resistance or glucose intolerance (theinability to properly use insulin or blood sugar), a chronicprothrombotic state (e.g., characterized by high fibrinogen orplasminogen activator inhibitor levels in the blood), and a chronicproinflammatory state (e.g., characterized by higher than normal levelsof high-sensitivity C-reactive protein in the blood). People withMetabolic Syndrome are at increased risk of coronary heart disease,other diseases related to plaque buildups in artery walls (e.g., strokeand peripheral vascular disease) and type 2 diabetes. The underlyingcauses of metabolic syndrome are unclear. Genetics, diet, disruptedchronobiology, mood disorders and alcohol use or exposure to pollutantse.g., tobacco may be implicated in Metabolic Syndrome.

In some embodiments, the Metabolic Syndrome is associated withresistance to insulin-mediated glucose uptake, glucose intolerance,hyperinsulemia, increased LDL-cholesterol, increased VLDL, increasedtriglycerides, decreased HDL-cholesterol, increased plasminogenactivator inhibitor-I (PALI) levels and hypertension. In certainembodiments, Metabolic Syndrome is characterized by three or more of thefollowing criteria: (a) abdominal obesity: waist circumference >102 cmin men and >88 cm in women; (b) hypertriglyceridemia: 150 mg/dl (1.695mmol/1); (c) low HDL cholesterol: <40 mg/dl (1.036 mmol/1) in men and<50 mg/dl (1.295 mmol/1) in women; (d) high blood pressure: 130/85 mmHg; and (e) high fasting glucose: 110 mg/dl (>6.1 mmol/1).

In some embodiments, Metabolic Syndrome is characterized by diabetes,impaired glucose tolerance, impaired fasting glucose, or insulinresistance plus two or more of the following abnormalities: (a) highblood pressure: 160/90 mm Hg; (b) hyperlipidemia: triglycerideconcentration 150 mg/dl (1.695 mmol/1) and/or HDL cholesterol <35 mg/dl(0.9 mmol/1) in men and <39 mg/dl (1.0 mmol/1) in women; (c) centralobesity: waist-to-hip ratio of >0.90 in men or >0.85 in women or BMI >30kg/m²; and (d) microalbuminuria: urinary albumin excretion rate 20μg/min or an albumin to creatinine ratio 20 mg/g.

In certain embodiments, Metabolic Syndrome is characterized by three ormore of the following criteria: (a) triglycerides>150 mg/dl; (b)systolic blood pressure (BP) 130 mm Hg or diastolic blood pressure 85 mmHg or on antihypertensive treatment; (c) high-density lipoproteincholesterol <40 mg/dl; (d) fasting blood sugar (FBS)>110 mg/dl; and (e)body mass index >28.8 kg/m².

The compositions disclosed herein promote glucose uptake into cells.

Components of the Formulations of the Present Technology

Lipoic acid is a naturally occurring substance found in the R form,which is considered the active species. The S form of lipoic acid isproduced during chemical synthesis and while having some antioxidantproperties, is considered to be inactive and toxic. It can be producedfrom octanoic acid in cells and can become covalently attached to enzymesubunits. Lipoic acid is an essential cofactor for a variety ofdehydrogenase enzyme systems in the mitochondria and plays an importantrole in the regulation of insulin receptor activity, glucose uptake andmetabolism. Alpha Lipoic Acid (ALA) (1,2-dithione-3-pentanoic acid),known as dihydrolipoic acid (DHLA) in its reduced form, is asulfur-containing saturated fatty acid found in small amounts in redmeat, organ meats, spinach, broccoli, peas, Brussels sprouts, potatoes,yams, carrots, beets, and yeast. As a supplement, ALA is rapidlyabsorbed into the blood and the cells where it is reduced to DHLA byNADH or NADPH in most tissues. ALA is also synthesized de nova fromoctanoic acid in mitochondria. ALA occurs naturally in every cell of thebody and is an essential cofactor for the mitochondrial dehydrogenasecomplexes which regulate mammalian glucose homoeostasis.

The quaternary ammonium compound N-acetyl-L-carnitine is involved intransporting fatty acids into mitochondria so that they can be used as afuel for energy production. The acetyl group of N-acetyl-L-carnitine isused to form acetyl-CoA, an intermediary in the tricarboxylic acid (TCA)cycle that is used to generate energy from amino acids, fats, andcarbohydrates.

Coenzyme QIO, also known as ubiquinone, ubidecarenone, coenzyme Q(CoQIO, CoQ, Q10, or Q), has been shown to improve brain and muscleenergy metabolism in the electron transfer process and plays a role inthe permeability of the inner mitochondrial membrane. (Alfredo Bianchiet al., Vitamins and Hormones, Vol. 69 2004 p. 297-312). The twoelectron oxidation of NADH in the electron transport chain is performedby coenzyme Q10 located in the mitochondrion membrane. The subsequentreduction of CoQ10 generates a proton gradient that strengthens theelectron transfer chain, increasing not only the fat synthesis pathwaythrough lipogenesis, but also increasing the overall efficiency of theTCA cycle.

L-Glutamine is an alternate carbon source to glucose in the TCA cycle,replenishing the cycle by oxidation of fatty acids to acetyl CoA.

L-arginine feeds into signal pathways of the TCA cycle. It is theimmediate precursor of nitric oxide, an important cellular signalingmolecule involved in many physiological and pathological processes.L-arginine is a powerful vasodilator, decreasing blood pressure andreducing atherosclerotic plaque that is associated with MetabolicSyndrome.

In some embodiments, the vitamin D compound formulations of the presenttechnology comprise molecules such as 5-HT (5-hydroxytryptamine),tryptophan, serotonin, melatonin, their analogues and precursors as wellas inhibitors of their degradation so as to promote sleep in thesubject.

Vitamin D enhances the absorption of essential minerals such as calcium,phosphorous, iron, magnesium and zinc. Supplementation of vitamin Dduring rest allows the body to restore and heal naturally.

Methylsulfonylmethane is a natural source of biologically active sulfur.

Chamomile powder derived from flowers, seeds, and pollen may be includedin the vitamin D compound formulations of the present technology.

The hormone melatonin is produced by the pineal gland among othertissues and is useful in the phase-dependent treatment of delayed sleepphase syndrome See Mundey K, Benloucif S, Harsanyi K, Dubocovich ML, andZee PC (2005). Phase-dependent treatment of delayed sleep phase syndromewith melatonin. Sleep 28:1271-1278.

5-HTP (5-hydroxytryptophan), also known as oxitriptan, is a naturallyoccurring amino acid, chemical precursor as well as a metabolicintermediate in the biosynthesis of the neurotransmitter serotonin.

Valerian is an extract of the roots of valerian plant (Valerianaofficinalis, Caprifoliaceae) (available at a number of commercialvendors including, but not limited to, Parchem, New Rochelle, N.Y., USAand Nutra Green, China).

Formulations of the Present Technology

The present disclosure provides therapeutic formulations intended toimprove glucose uptake and the oxidation of nutrients in mammals,including humans. The thiol-containing alkyl fatty acid formulations ofthe present technology comprise a thiol-containing alkyl fatty acid,co-formulated with anti-oxidants, nitrogen donors, TCA cycleintermediates and herbal extracts and excipients. Additionally oralternatively, in some embodiments, thiol-containing alkyl fatty acidsare co-formulated with naturally-occurring or synthetic antioxidants,amino acids, peptides, protease inhibitors, trace elements, botanicalextracts, and carbohydrates.

In one aspect, the present technology provides thiol-containing alkylfatty acid formulations comprising a thiol-containing alkyl fatty acid,a N-acetyl-L-carnitine derivative, an antioxidant, and at least oneamino acid. In certain embodiments, the thiol-containing alkyl fattyacid is lipoic acid or alpha-lipoic acid. In certain embodiments, thelipoic acid or alpha-lipoic acid is an R isomer, S isomer, or a mixtureof both Rand S isomers.

Additionally or alternatively, in some embodiments, theN-acetyl-L-carnitine derivative is L-carnitine. In certain embodiments,the amino acid is selected from among L-Glutamine and L-Arginine.

In some embodiments, the thiol-containing alkyl fatty acid is a compoundhaving the general formula:

or derivatives, congeners, metallic coordination complexes and saltsthereof,

wherein R₃ is an alkyl defined as C_(n)H_(2n+2), an alkenyl defined asC_(n)H_(2n), and/or an alkynyl defined as C_(n), where n is 1 to 18;

wherein R₁ and/or R₂ is aryl and/or aromatic;

wherein R₄, is an alkyl, alkenyl, alkynyl, aryl, —COOH, —OH, or —NH₂;and

wherein R₁, R₂, R₃, and/or R₄ is optionally phosphorylated.

In some embodiments, the N-acetyl-L-carnitine derivative is a compoundhaving the general formula:

or derivatives, congeners, metallic coordination complexes and saltsthereof,

wherein R is N or sulfonium;

and wherein R₁ is H; an alkyl defined as C_(n)H_(2n+2), an alkenyldefined as C_(n)H_(2n), and/or an alkynyl defined as Cn, where n is 1 to18; aryl; aromatic; —COOH; or —NH₂. In certain embodiments, thederivative is propionyl L-carnitine.

Additionally or alternatively, in some embodiments, the antioxidant isCoQ10, quercetin, or other plant-derived flavonoids.

Additionally or alternatively, in some embodiments, the amino acids areselected from among N-acetyl-L-carnitine and/or derivatives thereof,propionyl L-carnitine, L-carnitine, L-tartarate, L-glutamine, L-theanineand analogs thereof, L-arginine, and analogs thereof, and “essential”amino acids, precursors, analogs, and metabolites thereof.

In any of the above embodiments, the thiol-containing alkyl fatty acidformulations further comprise melatonin, peptides, protease inhibitors,trace elements, botanical extracts, and carbohydrates. In someembodiments, the proteins or peptides include serratiopeptidase, and/oranalogs, congeners, and mimetics thereof. In certain embodiments, thetrace elements include indium sulfate. In some embodiments, the plantextracts include eggplant extract, resveratrol-containing extracts suchas grape and grape seed extracts, guarana extract, purified or syntheticwhole-plant cannabis extract, vanilla extract, cinnamon, nutmeg, cloves,and turmeric. These botanical extracts are commercially available at anumber of suppliers (e.g., Parchem, New Rochelle, N.Y., USA; StarwestBotanicals, Sacramento, Calif.). It is anticipated that botanicalextracts obtained from any supplier will be useful in the preparation ofthe formulations of the present technology. In some embodiments, theprotease inhibitor is soybean trypsin inhibitor, leupeptin, pepstatin,or analogues, derivatives, mimetics and/or a combination thereof. Insome embodiments, the carbohydrates are selected from amongbeta-glucans, fiber extracts, pulps, simple and complex sugars andanalogs thereof. In certain embodiments, the beta-glucan is -1,3-glucanor -1,6-glucan.

In some embodiments, the thiol-containing alkyl fatty acid formulationcomprises a wt % from about 1% to about 5%, from about 5% to about 20%,from about 20% to about 30%, or at least about 31%, 32%, 33%, 34%, 35%,36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, or 65% of a thiol-containing alkyl fatty acid, such as lipoic acidor alpha-lipoic acid, relative to all of the components of theformulation.

In some embodiments, the thiol-containing alkyl fatty acid formulationcomprises a wt % from about 1% to about 5%, from about 5% to about 20%,from about 20% to about 30%, or at least about 31%, 32%, 33%, 34%, 35%,36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, or 65% of an N-acetyl L-camitine derivative, such as L-camitine,relative to all of the components of the formulation.

In some embodiments, the thiol-containing alkyl fatty acid formulationcomprises a wt % from about 1% to about 5%, from about 5% to about 20%,from about 20% to about 30%, or at least about 31%, 32%, 33%, 34%, 35%,36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, or 65% of an antioxidant, such as CoQ10, relative to all of thecomponents of the formulation.

In some embodiments, the thiol-containing alkyl fatty acid formulationcomprises a wt % from about 1% to about 5%, from about 5% to about 20%,from about 20% to about 30%, or at least about 31%, 32%, 33%, 34%, 35%,36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%,50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%,64%, or 65% of an amino acid, such as L-Glutamine or L-Arginine,relative to all of the components of the formulation.

In some embodiments, the ratio of the thiol-containing alkyl fatty acid(e.g., alpha-lipoic acid) to the N-acetyl L-camitine derivative (e.g.,L-camitine) in the thiol-containing alkyl fatty acid formulation of thepresent technology is about 2:1, 1.9:1, 1.8:1, 1.6:1, 1.5:1, 1.4:1,1.3:1, 1.2:1 or 1.1:1.

In some embodiments, the ratio of the thiol-containing alkyl fatty acid(e.g., alpha-lipoic acid) to the antioxidant (e.g., CoQIO) in thethiol-containing alkyl fatty acid formulation of the present technologyis about 8:1, 7:1, 6.5:1, 6:1, 5.5:1, 5:1, 4:1 or 3:1.

In some embodiments, the ratio of the thiol-containing alkyl fatty acid(e.g., alpha-lipoic acid) to the amino acid (e.g., L-Glutamine orL-Arginine) in the thiol-containing alkyl fatty acid formulation of thepresent technology is about 1:1, 1:1.1, 1:1.25, 1:1.3, 1:1.4, 1:1.5,1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5,1:2.6, 1:2.7, 1:2.8, 1:2.9, or 1:3.

Additionally or alternatively, the thiol-containing alkyl fatty acid,the N-acetyl-L-carnitine derivative, the antioxidant, or the at leastone amino acid is present in a thiol-containing alkyl fatty acidformulation useful in accordance with methods described herein in anamount of from about 50 mg to about 100 mg, or from about 100 mg toabout 150 mg, or from about 150 mg to about 200 mg, or from about 200 mgto about 250 mg, or from about 300 mg to about 350 mg, or from about 400mg to about 450 mg, or from about 450 mg to about 500 mg, or from about500 mg to about 600 mg, or from about 600 mg to about 800 mg, or fromabout 800 mg to about 1,000 mg, or from about 1,000 mg to about 1,200mg, or from about 1,200 mg to about 1,400 mg, or from about 1,400 mg toabout 1,600 mg, or from about 1,600 mg to about 1,800 mg, or from about1,800 mg to about 2,000 mg, or from about 2,000 mg to about 2,200 mg, orfrom about 2,200 mg to about 2,500 mg.

Also provided herein are vitamin D compound formulations comprisingamino acids, peptides, nucleic acids, carbohydrates, vitamins, otherorganic molecules and trace elements and plant extracts.

In one aspect, the present technology provides vitamin D compoundformulations comprising a vitamin D compound, a sulfur donor and one ormore sedatives or sleep promoting agents. In some embodiments, thevitamin D compound is cholecalciferol (D3), ergocalciferol (D2) or othervitamin D analogs. Additionally or alternatively, in some embodiments,the sulfur donor is methylsulfonylmethane.

Additionally or alternatively, in some embodiments, the sedatives orsleep promoting agents are one or more of melatonin, chamomile extract,valerian extract, 5-hydroxytryptamine (5-HT), SHTP(L-5-hydroxytryptophan), L-theanine, D-phenylalanine, L-phenylalanine,or a mixture of D- and L-phenylalanine.

Additionally or alternatively, in some embodiments, the vitamin Dcompound formulations of the present technology further comprisepeptides, proteins, trace elements, plant extracts, protease inhibitors,and carbohydrates. In some embodiments, the proteins or peptides includeserratiopeptidase, and/or analogs, congeners, and mimetics thereof. Incertain embodiments, the trace elements include indium sulfate. In someembodiments, the plant extracts include eggplant extract,resveratrol-containing extracts such as grape and grape seed extracts,guarana extract, purified or synthetic whole-plant cannabis extract,vanilla extract, cinnamon, nutmeg, cloves, and turmeric. These botanicalextracts are commercially available at a number of suppliers (e.g.,Parchem, New Rochelle, N.Y., USA; Starwest Botanicals, Sacramento,Calif.). It is anticipated that botanical extracts obtained from anysupplier will be useful in the preparation of the formulations of thepresent technology. In some embodiments, the protease inhibitor issoybean trypsin inhibitor, leupeptin, pepstatin, or analogues,derivatives, mimetics and/or a combination thereof. In some embodiments,the carbohydrates are selected from among beta-glucans, fiber extracts,pulps, simple and complex sugars and analogs thereof. In certainembodiments, the beta-glucan is -1,3-glucan or -1,6-glucan.

Additionally or alternatively, the sulfur donor, or the one or moresedatives or sleep promoting agents is present in a vitamin D compoundformulation useful in accordance with methods described herein in anamount of from about 1 mg to about 10 mg, from about 10 mg to about 50mg, from about 50 mg to about 100 mg, or from about 100 mg to about 150mg, or from about 150 mg to about 200 mg, or from about 200 mg to about250 mg, or from about 300 mg to about 350 mg, or from about 400 mg toabout 450 mg, or from about 450 mg to about 500 mg, or from about 500 mgto about 600 mg, or from about 600 mg to about 800 mg, or from about 800mg to about 1,000 mg.

In some embodiments, the vitamin D compound formulations disclosedherein restore or enhance the body's own ability to adjust hormonelevels and metabolic function during sleep. In certain embodiments, thesleep inducing and enhancing components are formulated in a controlledrelease formulation such that (1) the sedating and sleep inducingcomponents such as chamomile and melatonin are released immediately, (2)the sleep maintaining components such as 5-HTP are slowly released overtime, maintaining a consistent blood level through the sleep period and(3) the REM promoting components, such as valerian is delayed releasedto initial the REM sleep cycle appropriately. In yet a further exampleembodiment, a bronchial dilator is combined with the sleep inducing andenhancing components to prevent or reduce sleep apnea.

In any of the above embodiments, the thiol-containing alkyl fatty acidformulations and vitamin D compound formulations of the presenttechnology may include pharmaceutically acceptable carriers andexcipients.

In any of the above embodiments, at least two, at least three, or atleast four or more of the individual components of the thiol-containingalkyl fatty acid formulations and vitamin D compound formulations of thepresent technology have overlapping oral bioavailability.

Additionally or alternatively, the thiol-containing alkyl fatty acidformulations and vitamin D compound formulations of the presenttechnology may be provided as an oral solid medicament in the form oftablets, capsules, powders, nanoparticles, or nano-emulsions forimmediate or sustained release. The thiol-containing alkyl fatty acidformulations and vitamin D compound formulations of the presenttechnology may comprise a plurality of crystalline structures. Thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations can also contain additional ingredients or encapsulationsthat establish the release profile of each or the collectiveingredient(s) for instantaneous release, or sustained release. In afurther embodiment, the oral solid dosage form can be enterically coatedso as to prevent gastric irritation in the subject. Alternatively, thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations of the present technology can be either fully dissolved orprepared as an emulsion. Additionally or alternatively, thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations of the present technology may be provided as a liquid.

In another aspect, the thiol-containing alkyl fatty acid formulationsand vitamin D compound formulations of the present technology compriseacid analogs of the components disclosed herein, which are buffered byion-pairing salts to minimize rapid degradation, increase absorption,and to minimize stomach irritation or acid reflux.

The present disclosure thus provides a diurnal regimen comprising athiol-containing alkyl fatty acid formulation and a vitamin D compoundformulation.

Therapeutic Methods

The following discussion is presented by way of example only, and is notintended to be limiting. The present technology also provides methods ofusing the formulations disclosed herein to prevent or treat diabetes,pre-diabetes, Metabolic Syndrome.

One aspect of the present technology includes methods of treatingpre-diabetes, diabetes or Metabolic Syndrome in a subject diagnosed ashaving, suspected as having, or at risk of having pre-diabetes, diabetesor Metabolic Syndrome. In therapeutic applications, formulations of thepresent technology are administered to a subject suspected of, oralready suffering from such a disease (such as, e.g., subjects with IFGand/or IGT, insulin resistance or elevated glycated hemoglobin levels(HbA1c) or a subject diagnosed with pre-diabetes, diabetes or MetabolicSyndrome), in an amount sufficient to cure, or at least partiallyarrest, the symptoms of the disease, including its complications andintermediate pathological phenotypes in development of the disease.

Subjects suffering from pre-diabetes, diabetes or Metabolic Syndrome canbe identified by any or a combination of diagnostic or prognostic assaysknown in the art (e.g., fasting glucose or glucose tolerance, liver andpancreas laboratory panels and levels of glycosylated hemoglobin). Inaddition to height, weight, blood pressure, cardiovascular and pulmonaryfunctions, body mass index (BMI) has emerged as a likely indicator ofmetabolic dysfunction (seejournals.plos.org/plosmedicine/article?id=10.1371/journal.pmed.1001230#). Body waist size may correlate with the risk of type II diabetes.

For example, typical symptoms of pre-diabetes, diabetes or MetabolicSyndrome include, but are not limited to, body weight, fastingglucose/insulin/free fatty acid, glucose tolerance (OGTT), muscleinsulin sensitivity, markers of insulin signaling (e.g., Akt-P, IRS-P),serum triglyceride levels, HDL and LDL cholesterol levels, bloodpressure, serum fibrinogen or plasminogen activator inhibitor levels,levels of C-reactive protein, insulin resistance, glucose intolerance,elevated levels of glycosylated blood proteins (e.g., hemoglobin).

In some embodiments, pre-diabetic, diabetic or Metabolic Syndromesubjects treated with the formulation of the present technology willshow amelioration or elimination of one or more of the followingsymptoms: body weight, fasting glucose/insulin/free fatty acid, glucosetolerance (OGTT), muscle insulin sensitivity, markers of insulinsignaling (e.g., Akt-P, IRS-P), serum triglyceride levels, HDL and LDLcholesterol levels, blood pressure, serum fibrinogen or plasminogenactivator inhibitor levels, levels of C-reactive protein, insulinresistance, glucose intolerance, elevated levels of glycosylated bloodproteins (e.g., hemoglobin).

In some embodiments of the methods of the present technology, MetabolicSyndrome is treated through improvement of lipid metabolism compared tothe subject's lipid metabolism before being administered theformulations disclosed herein. In one embodiment, the improvement inlipid metabolism is reduction of blood triglyceride level compared tothe subject's blood triglyceride level before being administered theformulations of the present technology. In another embodiment, theimprovement in lipid metabolism is improvement of blood HDL/LDLcholesterol ratio compared to the subject's blood HDL/LDL cholesterolbefore being administered the formulations of the present technology.For instance, a subject may exhibit at least about 5%, at least about10%, at least about 20%, or at least about 50% reduction in body weightcompared to the subject prior to receiving the formulations of thepresent technology. In one embodiment, a subject may exhibit at leastabout 5%, at least about 10%, at least about 20%, or at least about 50%reduction in HDL cholesterol and/or at least about 5%, at least about10%, at least about 20%, or at least about 50% increase in LDLcholesterol compared to the subject prior to receiving the formulationsof the present technology. In one embodiment, a subject may exhibit atleast about 5%, at least about 10%, at least about 20%, or at leastabout 50% reduction in serum triglycerides compared to the subject priorto receiving the formulations of the present technology. In oneembodiment, a subject may exhibit at least about 5%, at least about 10%,at least about 20%, or at least about 50% improvement in oral glucosetolerance (OGTT) compared to the subject prior to receiving theformulations of the present technology.

Additionally or alternatively, in some embodiments of the methods of thepresent technology, Metabolic Syndrome is treated by reducing bloodsugar level compared to the subject's blood sugar level before beingadministered the formulations of the present technology. In otherembodiments, Metabolic Syndrome is treated by a reduction in body weightcompared to the subject's body weight before being administered theformulations of the present technology.

In some cases, the insulin resistance may be due to a high fat diet or,more generally, over-nutrition. The formulations of the presenttechnology are useful in treating diabetic, pre-diabetic or obeseinsulin resistant, non-diabetic patients. Insulin resistance occurs whenthe body does not respond to the insulin made by the pancreas andglucose is less able to enter the cells. Subjects with insulinresistance may or may not go on to develop type 2 diabetes. In someembodiments, administration of an effective amount of the formulationsof the present technology improves at least one sign or symptom ofinsulin resistance in the subject, e.g., body weight, fastingglucose/insulin/free fatty acid, glucose tolerance (OGTT), in vitromuscle insulin sensitivity, markers of insulin signaling (e.g., Akt-P,IRS-P), mitochondrial function (e.g., respiration), mitochondrial enzymeactivity and the like.

Also provided are methods for treating a disease or conditioncharacterized by elevated glycated hemoglobin levels (HbA1c) in asubject in need thereof comprising administering to the subject aneffective amount of the formulations of the present technology to thesubject. In some embodiments, administration of an effective amount ofthe formulations of the present technology causes a reduction in theglycated hemoglobin levels (HbA1c) of the subject. In some embodiments,the disease or condition characterized by elevated glycated hemoglobinlevels (HbA1c) is pre-diabetes, diabetes, Metabolic Syndrome, insulinresistance, glucose intolerance or cardiovascular disease, and stroke.In some embodiments, administration of an effective amount of theformulations of the present technology causes a reduction in theglycated hemoglobin levels (HbA1c) of the subject by at least 3%, by atleast 4%, by at least 5%, by at least 6%, by at least 8%, or by at least10% compared to that observed in the subject prior to treatment.

Prophylactic Methods

In one aspect, the present technology provides a method for preventingor delaying the onset of pre-diabetes, diabetes or Metabolic Syndrome orsymptoms of pre-diabetes, diabetes or Metabolic Syndrome in a subject atrisk of having pre-diabetes, diabetes or Metabolic Syndrome. In someembodiments, the subject may exhibit IFG and/or IGT, insulin resistance,and/or elevated glycated hemoglobin levels (HbA1c).

Subjects suffering from pre-diabetes, diabetes or Metabolic Syndrome canbe identified by, e.g., any or a combination of diagnostic or prognosticassays known in the art. In prophylactic applications, the formulationsof the present technology are administered to a subject susceptible to,or otherwise at risk of a disease or condition such as e.g.,pre-diabetes, diabetes or Metabolic Syndrome, in an amount sufficient toeliminate or reduce the risk, or delay the onset of the disease,including biochemical, histologic and/or behavioral symptoms of thedisease, its complications and intermediate pathological phenotypespresenting during development of the disease. Administration of aprophylactic formulation of the present technology can occur prior tothe manifestation of symptoms characteristic of the disease or disorder,such that the disease or disorder is prevented or, alternatively,delayed in its progress 10n.

For therapeutic and/or prophylactic applications, one or moreformulations of the present technology are administered to the subject.In some embodiments, the thiol-containing alkyl fatty acid formulationsand vitamin D compound formulations are administered one, two, three,four, or five times per day. In some embodiments, the thiol-containingalkyl fatty acid formulations and vitamin D compound formulations areadministered more than five times per day. Additionally oralternatively, in some embodiments, the thiol-containing alkyl fattyacid formulations and vitamin D compound formulations are administeredevery day, every other day, every third day, every fourth day, everyfifth day, or every sixth day. In some embodiments, the thiol-containingalkyl fatty acid formulations and vitamin D compound formulations areadministered weekly, bi-weekly, tri-weekly, or monthly. In someembodiments, the thiol-containing alkyl fatty acid formulations andvitamin D compound formulations are administered for a period of one,two, three, four, or five weeks. In some embodiments, thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations are administered for six weeks or more. In someembodiments, the thiol-containing alkyl fatty acid formulations andvitamin D compound formulations are administered for twelve weeks ormore. In some embodiments, the thiol-containing alkyl fatty acidformulations and vitamin D compound formulations are administered for aperiod of less than one year. In some embodiments, the thiol-containingalkyl fatty acid formulations and vitamin D compound formulations areadministered for a period of more than one year. In some embodiments,the thiol-containing alkyl fatty acid formulations and vitamin Dcompound formulations of the present technology are administered orally,topically, intranasally, systemically, intravenously, subcutaneously,intraperitoneally, intradermally, iontophoretically, transmucosally, orintramuscularly.

In some embodiments of the methods of the present technology, thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations of the present technology are administered daily for 1 weekor more. In some embodiments of the methods of the present technology,the thiol-containing alkyl fatty acid formulations and vitamin Dcompound formulations of the present technology are administered dailyfor 2 weeks or more. In some embodiments of the methods of the presenttechnology, the thiol-containing alkyl fatty acid formulations andvitamin D compound formulations of the present technology areadministered daily for 3 weeks or more. In some embodiments of themethods of the present technology, the thiol-containing alkyl fatty acidformulations and vitamin D compound formulations of the presenttechnology are administered daily for 4 weeks or more. In someembodiments of the methods of the present technology, thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations of the present technology are administered daily for 6weeks or more. In some embodiments of the methods of the presenttechnology, the thiol-containing alkyl fatty acid formulations andvitamin D compound formulations of the present technology areadministered daily for 12 weeks or more.

In some embodiments, treatment with the thiol-containing alkyl fattyacid formulations and vitamin D compound formulations of the presenttechnology will prevent or delay the onset of one or more of thefollowing symptoms: increased body weight, elevated serum triglyceridelevels, decreased HDL and/or increased LDL cholesterol levels, highblood pressure, insulin resistance, glucose intolerance, elevated levelsof glycosylated blood proteins (e.g., hemoglobin), alterations in one ormore of the following compared to healthy control subjects—fastingglucose/insulin/free fatty acid, glucose tolerance (OGTT), muscleinsulin sensitivity, markers of insulin signaling (e.g., Akt-P, IRS-P),serum fibrinogen or plasminogen activator inhibitor levels, levels ofC-reactive protein, etc.

EXAMPLES

The present technology is further illustrated by the following examples,which should not be construed as limiting in any way. For each of theexamples below, any formulation as described herein could be used.

Example 1: Methods of Treatment Using Formulation a and Formulation B

This Example demonstrates that the formulations of the presenttechnology are useful in treating pre-diabetes, diabetes or MetabolicSyndrome in human subjects. Many of the individual components of thedisclosed formulations are commercially available at a number ofsuppliers. The suppliers disclosed herein are intended to be exemplary.It is anticipated that there are multiple commercial sources for theindividual components of the compositions of the present technologyuseful in the preparation of the formulations of the present technology.

Compositions

Formulation A is an example of a thiol-containing alkyl fatty acidformulation for the treatment of pre-diabetes, diabetes, MetabolicSyndrome or a disease characterized by abnormal glucose metabolism.

The components of Formulation A are lipoic acid, L-camitine, CoQ10,L-glutamine and L-arginine. One advantage of this formulation is theavailability of arginine as a nitric oxide donor in the presence ofglutamine, a carbon source for mitochondrial function and aneurotransmitter precursor.

Formulation A Ingredient Amount Alpha-lipoic acid (Changshu WanxingChemical Co. Ltd., 600 mg China) L-Camitine (Kaiyuan Hengtai ChemicalCo., Ltd. China) 400 mg CoQI0 (Inner Mongolia Kingdomway PharmaceuticalsCo., 100 mg Ltd., China) L-Glutamine (Kyowa Hakko Kogyo Co., Japan) 750mg L-Arginine (Shine Star (Hubei) Biological Engineering Co., 1500 mgLtd., China)

The components of Formulation A are both allosteric regulators(prosthetic groups for selected enzymes, genetic and epigeneticstimulators of activity), and regulators of transport molecules thatconvey glucose, lipids, and fatty acids to the mitochondria. Withoutwishing to be bound by theory, the net result may be an increasedability to transport glucose out of the blood and into tissuesindependent of insulin levels.

Further, over the course of treatment, levels of glucose transporterproteins may be increased;

Formulation B is an example of a vitamin D compound formulation for thetreatment of pre-diabetes, diabetes, Metabolic Syndrome or a diseasecharacterized by abnormal glucose metabolism.

Formulation B Ingredient Amount Vitamin D (Prinova Solutions, CarolStream, IL, USA) 1000 IU Methylsulfonylmethane (e.g., FoodchemInternational 400 mg Corporation, China) 5-HTP (Gryffonia simplicfoliaseed) (Jiangsu Swellxin Bio- 50 mg pharm Pty. Ltd., China) Melatonin(Wuhan Yuancheng Technology Development 3 mg Co., Ltd., China) Valerianroot powder (e.g., Parchem, New Rochelle, New 600 mg York, USA)Chamomile powder (isolated from flowers, seeds, pollen) 500 mg (e.g.,Parchem, New Rochelle, New York, USA) L-Theanine (Sichuan TongshengAmino Acid Co., Ltd., 400 mg China)

Methods

Three male patients received a daily dose of Formulation A andFormulation B for 90 days. BMI, heart rate, blood pressure, and HbA1clevels were measured prior to treatment (Day 0) and 90 days postadministration of Formulation A and Formulation B. See Tables 1 and 2.

TABLE 1 Heart Rate Blood Pressure HbAlc 1 Patient Age Day 0 Day 90 Day 0Day 90 Day 0 Day 90 1 76 92 70 112/75 120/80 7.0 6.7 2 90 70 66 155/75150/70 6.1 5.8 3 47 N/a N/a 135/90 128/86 5.7 5.8

HbA1c levels can indicate people with pre-diabetes ordiabetes—Specifically, the HbA1c in normal subjects is below 6.0%,whereas the HbA1c is 6.0-6.4% in pre-diabetic subjects and 6.5% indiabetic subjects. As shown in Table 1, patients 1 and 2 exhibited anobservable reduction in HbA1c after receiving a daily dose ofFormulations A and B for 90 days. Further, patient 1 showed a reductionin the severity of pre-diabetes whereas patient 2, which initially hadpre-diabetic HbA1c levels, exhibited normal HbA1c level post-treatment.HbA1c levels can also be used to predict the estimated average bloodglucose (mg/dL). See FIGS. 1A and 1B. The estimated average bloodglucose for patient 1 and patient 2 post-treatment are 146 mg/dL and 120mg/dL respectively. Both patient 1 and patient 2 showed a 8 mg/dLdecrease in the estimated average blood glucose at the end of the 90 daytreatment window, which corresponds to a 5.2% and 6.2% decrease inestimated average blood glucose in patient 1 and patient 2 respectively.

Table 1 also shows that administration of Formulations A and B improvedheart rate and blood pressure in some patients.

TABLE 2 Weight BMI Patient Height Day 0 Day 90 Day 0 Day 90 1 67.0 164159 25.7 24.1 2 68.5 166.0 168.5 24.9 25.2 3 72.0 218 213 29.6 28.9

TABLE 3 BMI Weight Status Below 18.5 Underweight 18.5-24.9 Normal orHealthy Weight 25.0-29.9 Overweight 30.0 and Above Obese

As shown in Tables 2 and 3, patient 1 showed about a 7% decrease in BMIand a change in weight status from “overweight” to a “healthy weight” atthe end of the 90 day treatment window. Table 2 shows that patient 3exhibited a mild improvement in BMI at the end of the 90 day treatmentwindow. No adverse events were reported. One out of the three patientsshowed a dramatic improvement in restorative sleep.

These results demonstrate that the thiol-containing alkyl fatty acidformulations and vitamin D compound formulations of the presenttechnology are effective in reducing elevated levels of glycatedhemoglobin (HbA1c) in mammalian subjects. Accordingly, thethiol-containing alkyl fatty acid formulations and vitamin D compoundformulations disclosed herein are useful for treating pre-diabetes,diabetes, Metabolic Syndrome or a condition associated with abnormalglucose metabolism in a subject in need thereof.

Example 2: Treatment of Diabetes as Measured by Reduction in HbA1c

This Example will show that the formulations of the present technologyare useful in the treatment of diabetes.

Methods

Animals: Male mice (ddY, 5 weeks old) are kept in an experimental animalroom for 7 days with free access to food and water. Neonatalstreptozotocin-induced diabetic mice (NSZ) are produced by subcutaneousinjection of streptozotocin (STZ), 90 mg/kg body weight, which isdissolved in citrate buffer (pH 4.5). Non-STZ injected mice are used asa control. Five weeks after injection of STZ, the blood glucose levelsof all the mice are determined. Mice with a blood glucose level above250 mg/dl are considered to be diabetic and are used in the study.

Determination of Glucose and Hemoglobin Ale (HbAlc): To identifydiabetic mice, blood samples are withdrawn from the cavernous sinus witha capillary in STZ treated mice and blood glucose levels are determinedby the glucose oxidase method.

HbA1c levels are measured from blood samples from test subjects by animmunoassay (see, e.g., DCA-2000 System, Bayer-Sankyo Co., Ltd. Tokyo,Japan).

Treatment: The mice are divided into groups and treated according toTables 4-7 (see Example 1 for composition of Formulation A andFormulation B):

TABLE 4 Diabetic Mice First Treatment Group Treatment 1 Untreated 2Formulation A 3 Alpha-lipoic acid (Changshu Wanxing Chemical Co. Ltd.,China) 4 L-Carnitine (Kaiyuan Hengtai Chemical Co., Ltd. China) 5 CoQ10(Inner Mongolia Kingdomway Pharmaceuticals Co., Ltd., China) 6L-Glutamine (Kyowa Hakko Kogyo Co., Japan) 7 L-Arginine (Shine Star(Hubei) Biological Engineering Co., Ltd., China) 8 Vitamin D (PrinovaSolutions, Carol Stream, IL, USA) 9 Methylsulfonylmethane (e.g.,Foodchem International Corporation, China) 10 5-HTP (Gryffoniasimplicfolia seed) (Jiangsu Swellxin Bio- pharm Pty. Ltd., China) 11Melatonin (Wuhan Yuancheng Technology Development Co., Ltd., China) 12Valerian root powder (e.g., Parchem, New Rochelle, New York, USA) 13Chamomile powder (isolated from flowers, seeds, pollen) (e.g., Parchem,New Rochelle, New York, USA) 14 L-Theanine (Sichuan Tongsheng Amino AcidCo., Ltd., China)

TABLE 5 Control Mice First Treatment Group Treatment 15 Untreated 16Formulation A 17 Alpha-lipoic acid (Changshu Wanxing Chemical Co. Ltd.,China) 18 L-Carnitine (Kaiyuan Hengtai Chemical Co., Ltd. China) 19CoQ10 (Inner Mongolia Kingdomway Pharmaceuticals Co., Ltd., China) 20L-Glutamine (Kyowa Hakko Kogyo Co., Japan) 21 L-Arginine (Shine Star(Hubei) Biological Engineering Co., Ltd., China) 22 Vitamin D (PrinovaSolutions, Carol Stream, IL, USA) 23 Methylsulfonylmethane (e.g.,Foodchem International Corporation, China) 24 5-HTP (Gryffoniasimplicfolia seed) (Jiangsu Swellxin Bio- pharm Pty. Ltd., China) 25Melatonin (Wuhan Yuancheng Technology Development Co., Ltd., China) 26Valerian root powder (e.g., Parchem, New Rochelle, New York, USA) 27Chamomile powder (isolated from flowers, seeds, pollen) (e.g., Parchem,New Rochelle, New York, USA) 28 L-Theanine (Sichuan Tongsheng Amino AcidCo., Ltd., China)

TABLE 6 Diabetic Mice Second Treatment Group Treatment 1 Untreated 2Formulation B 3 Alpha-lipoic acid (Changshu Wanxing Chemical Co. Ltd.,China) 4 L-Carnitine (Kaiyuan Hengtai Chemical Co., Ltd. China) 5 CoQ10(Inner Mongolia Kingdomway Pharmaceuticals Co., Ltd., China) 6L-Glutamine (Kyowa Hakko Kogyo Co., Japan) 7 L-Arginine (Shine Star(Hubei) Biological Engineering Co., Ltd., China) 8 Vitamin D (PrinovaSolutions, Carol Stream, IL, USA) 9 Methylsulfonylmethane (e.g.,Foodchem International Corporation, China) 10 5-HTP (Gryffoniasimplicfolia seed) (Jiangsu Swellxin Bio- pharm Pty. Ltd., China) 11Melatonin (Wuhan Yuancheng Technology Development Co., Ltd., China) 12Valerian root powder (e.g., Parchem, New Rochelle, New York, USA) 13Chamomile powder (isolated from flowers, seeds, pollen) (e.g., Parchem,New Rochelle, New York, USA) 14 L-Theanine (Sichuan Tongsheng Amino AcidCo., Ltd., China)

TABLE 7 Control Mice Second Treatment Group Treatment 15 Untreated 16Formulation B 17 Alpha-lipoic acid (Changshu Wanxing Chemical Co. Ltd.,China) 18 L-Carnitine (Kaiyuan Hengtai Chemical Co., Ltd. China) 19CoQ10 (Inner Mongolia Kingdomway Pharmaceuticals Co., Ltd., China) 20L-Glutamine (Kyowa Hakko Kogyo Co., Japan) 21 L-Arginine (Shine Star(Hubei) Biological Engineering Co., Ltd., China) 22 Vitamin D (PrinovaSolutions, Carol Stream, IL, USA) 23 Methylsulfonylmethane (e.g.,Foodchem International Corporation, China) 24 5-HTP (Gryffoniasimplicfolia seed) (Jiangsu Swellxin Bio- pharm Pty. Ltd., China) 25Melatonin (Wuhan Yuancheng Technology Development Co., Ltd., China) 26Valerian root powder (e.g., Parchem, New Rochelle, New York, USA) 27Chamomile powder (isolated from flowers, seeds, pollen) (e.g., Parchem,New Rochelle, New York, USA) 28 L-Theanine (Sichuan Tongsheng Amino AcidCo., Ltd., China)

Each group includes 10 mice. All mice in Groups 3-14 and 17-28 aretreated with their respective compound twice daily by oraladministration (i.e., drinking water or gavage). All mice in Groups 2and 16 are treated daily with Formulation A once and Formulation B once.The dosages of each formulation or compound (disclosed in Example 1) areadjusted according to the proper ratio based on the average BMI of themice. Groups 3-14 and 17-28 are treated with an equivalent molar dose oftheir respective compounds based on the concentration of the respectivecompound (i.e., alpha-lipoic acid, L-carnitine, CoQ10, L-glutamine,L-arginine, vitamin D, methylsulfonylmethane, 5-HTP, melatonin, valerianroot powder, L-theanine and chamomile powder) in Formulation A andFormulation B administered in the Formulation A/Formulation B treatmentgroups. All mice are treated for 90 days. Blood is drawn from each mouseat 7 days post 1^(st) treatment, 14 days post 1^(st) treatment, 28 dayspost 1^(st) treatment, 56 days post 1^(st) treatment, 70 days post1^(st) treatment, and 90 days post 1^(st) treatment and the HbA1c levelis determined for each blood sample.

Results

It is anticipated that diabetic mice treated with Formulation A andFormulation B will show a reduction of HbA1c as compared to theuntreated diabetic control group. It is also anticipated that diabeticmice treated with Formulation A and Formulation B will show asynergistic effect by having a greater reduction of HbA1c, reduced sideeffects, and/or additional therapeutic benefits (including, but notlimited to reduced BMI, improved heart rate and blood pressure) ascompared to diabetic mice treated with the individual components ofFormulation A or Formulation B alone.

These results demonstrate that the formulations of the presenttechnology are effective in reducing elevated levels of glycatedhemoglobin (HbA1c) in mammalian subjects. These results will show thatFormulation A or Formulation B of the present technology are useful inmethods for treating diabetes in mammalian subjects.

Example 3: Prevention or Treatment of Insulin Resistance

This Example will show that the formulations of the present technologyare useful in the treatment of insulin resistance.

Methods

The fatty (fa/fa) Zucker rat, a model of diet-induced insulin resistanceis used. Young Zucker rats (˜3-4 weeks of age) and control wild-typerats are either untreated or administered Formulation A and FormulationB or their respective individual components as described in Tables 4-7in Example 2 for 90 days. Two parallel experiments are conducted to testthe prevention and treatment of insulin resistance.

Determination of Hemoglobin Alc (HbA1c): Blood is drawn from thesubjects at 7 days post 1^(st) treatment, 14 days post 1^(st) treatment,28 days post 1^(st) treatment, 56 days post 1^(st) treatment, 70 dayspost 1^(st) treatment, and 90 days post 1^(st) treatment and HbA 1clevel is determined for each blood sample. HbA1c is measured by animmunoassay (see, e.g., DCA-2000 System, Bayer-Sankyo Co., Ltd. Tokyo,Japan).

Results

It is anticipated that administration of Formulation A and Formulation Bwill attenuate or prevent the development of whole body and muscleinsulin resistance that normally develops in the fatty (fa/fa) Zuckerrat. Measured outcomes include body weight, fasting glucose/insulin/freefatty acid, glucose tolerance (OGTT), in vitro muscle insulinsensitivity (incubation), markers of insulin signaling (Akt-P, IRS-P),mitochondrial function studies on permeabilized fibers (respiration,H₂O₂ emission), and the like. A comparison is made between control ratsand fa/fa rats administrated Formulation A and Formulation B. Controlsinclude wild-type and fa/fa rats not administered Formulation A andFormulation B (i.e., the untreated rats). Successful prevention ofinsulin resistance by the administration of Formulation A andFormulation B is indicated by a reduction in one or more of the markersassociated with insulin resistance. It is anticipated that fa/fa ratstreated with Formulation A and Formulation B will show a reduction ofHbA1c as compared to the untreated fa/fa control group and that HbA1clevels of fa/fa rats treated with Formulation A and Formulation B willresemble those observed in the wild-type control rats.

It is anticipated that administration of Formulation A and Formulation Bwill ameliorate or reduce whole body and muscle insulin resistance thatnormally develops in these older fatty (fa/fa) Zucker rats. Measuredoutcomes include body weight, fasting glucose/insulin/free fatty acid,glucose tolerance (OGTT), in vitro muscle insulin sensitivity(incubation), markers of insulin signaling (Akt-P, IRS-P), mitochondrialfunction studies on permeabilized fibers (respiration, H₂O₂ emission),and mitochondrial enzyme activity. A comparison is made between controlrats and fa/fa rats administrated Formulation A and Formulation B.Controls include wild-type and fa/fa rats not administered Formulation Aand Formulation B (i.e., the untreated rats). Successful treatment ofinsulin resistance by the administration of Formulation A andFormulation Bis indicated by a reduction in one or more of the markersassociated with insulin resistance. It is anticipated that fa/fa ratstreated with Formulation A and Formulation B will show a reduction ofHbA1c as compared to the untreated fa/fa control group and that HbA1clevels of fa/fa rats treated with Formulation A and Formulation B willresemble those observed in the wild-type control rats.

It is also anticipated that Zucker rats administered Formulation A andFormulation B will show a synergistic effect by having a greaterreduction in one or more of the markers/symptoms associated with insulinresistance enumerated above and HbA1c levels as compared to Zucker ratstreated with the individual components of Formulation A and FormulationB alone.

These results demonstrate that the formulations of the presenttechnology are effective in reducing elevated levels of glycatedhemoglobin (HbA1c) in mammalian subjects. These results will show thatadministration of Formulation A and Formulation B of the presenttechnology are useful in methods for preventing and treating insulinresistance in mammalian subjects.

Example 4: Treatment of Metabolic Syndrome in Rats Fed a High Fat Diet

This Example will show that the formulations of the present technologyare useful in the treatment of Metabolic Syndrome.

Methods

In vivo model. A rat model of Metabolic Syndrome is established by acombination of 6-week high fat diet (HFD) and low-dose STZ (40 mg/kg)injection in Sprague-Dawley rats. Rats fed with normal chow (NRC) areused as a control. Rats are grouped and treated as described in Tables4-7 in Example 2. Metabolic Syndrome rats maintain the HFD and controlrats maintain the NRC diet throughout the 90 day treatment period.

Blood glucose. Blood glucose is tested by ACCU-CHEK® Advantage™ bloodglucose meter and test strips from Roche diagnostics GmbH (Mannheim,Germany).

Blood sample handling. Blood samples are collected by tail snip. In somecases when tail snip fails to provide sufficient volume, blood samplesare collected from retro-orbital sinus. Collected blood samples are setat room temperature for 30 min to allow clotting before beingcentrifuged at 1,200×g for 10 min. The supernatant is collected andre-centrifuged at 1,200×g for another 5 min to generate the serum. Serumsamples are aliquotted and stored at −20° C. until used in analysis.

Determination of Hemoglobin Alc (HbA1c): Blood is drawn from thesubjects at 7 days post 1^(st) treatment, 14 days post 1^(st) treatment,28 days post 1^(st) treatment, 56 days post 1^(st) treatment, 70 dayspost 1^(st) treatment, and 90 days post 1^(st) treatment and HbA1clevels are determined for each blood sample. HbA1c is measured by animmunoassay (see, e.g., DCA-2000 System, Bayer-Sankyo Co., Ltd. Tokyo,Japan).

Statistics: Statistic analysis oft-test and one-way ANOVA is performedby SPSS (Statistical Package for Social Science) analysis software.

Results

Effects of Administration of Formulation A and B on the body weight ofMetabolic Syndrome rats. It is anticipated that Metabolic Syndrome ratsadministered Formulation A and B will have reduced body weight ascompared to untreated Metabolic Syndrome rats. It is also anticipatedthat Metabolic Syndrome rats administered Formulation A and B will showa synergistic effect with respect to lower body weight as compared toMetabolic Syndrome rats treated with the individual components ofFormulation A and Formulation B alone. It is anticipated that MetabolicSyndrome rats administered Formulation A and B will exhibit body weightsthat resemble those observed in the wild-type control rats.

Effects of Administration of Formulation A and B on the blood glucose ofMetabolic Syndrome rats. It is anticipated that Metabolic Syndrome ratsadministered Formulation A and B will have reduced blood glucose ascompared to untreated Metabolic Syndrome rats. It is also anticipatedthat Metabolic Syndrome rats administered Formulation A and B will showa synergistic effect with respect to lower blood glucose levels ascompared to Metabolic Syndrome rats treated with the individualcomponents of Formulation A and Formulation B alone. It is anticipatedthat Metabolic Syndrome rats administered Formulation A and B willexhibit blood glucose levels that resemble those observed in thewild-type control rats.

Effects of Administration of Formulation A and B on HbA1c in MetabolicSyndrome rats. It is anticipated that Metabolic Syndrome ratsadministered Formulation A and B will have reduced HbA1c levels ascompared to the untreated Metabolic Syndrome control group. It is alsoanticipated that Metabolic Syndrome rats administered Formulation A andB will show a synergistic effect with respect to lower levels of HbA1cas compared to Metabolic Syndrome rats treated with the individualcomponents of Formulation A and Formulation B alone. It is anticipatedthat Metabolic Syndrome rats administered Formulation A and B willexhibit HbA1c levels that resemble those observed in the wild-typecontrol rats.

These results demonstrate that the formulations of the presenttechnology are effective in reducing elevated levels of glycatedhemoglobin (HbA1c) in mammalian subjects. These results will show thatadministration of Formulation A and Formulation B of the presenttechnology are useful in methods for treating Metabolic Syndrome inmammalian subjects.

EQUIVALENTS

The present technology is not to be limited in terms of the particularembodiments described in this application, which are intended as singleillustrations of individual aspects of the present technology. Manymodifications and variations of this present technology can be madewithout departing from its spirit and scope, as will be apparent tothose skilled in the art. Functionally equivalent methods andapparatuses within the scope of the present technology, in addition tothose enumerated herein, will be apparent to those skilled in the artfrom the foregoing descriptions. Such modifications and variations areintended to fall within the scope of the appended claims. The presenttechnology is to be limited only by the terms of the appended claims,along with the full scope of equivalents to which such claims areentitled. It is to be understood that this present technology is notlimited to particular methods, reagents, compounds compositions orbiological systems, which can, of course, vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to belimiting.

In addition, where features or aspects of the disclosure are describedin terms of Markush groups, those skilled in the art will recognize thatthe disclosure is also thereby described in terms of any individualmember or subgroup of members of the Markush group.

As will be understood by one skilled in the art, for any and allpurposes, particularly in terms of providing a written description, allranges disclosed herein also encompass any and all possible subrangesand combinations of subranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as “up to,” “at least,” “greater than,” “less than,” and the like,include the number recited and refer to ranges which can be subsequentlybroken down into subranges as discussed above. Finally, as will beunderstood by one skilled in the art, a range includes each individualmember. Thus, for example, a group having 1-3 cells refers to groupshaving 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers togroups having 1, 2, 3, 4, or 5 cells, and so forth. It is understood bythose of ordinary skill in the art that the labeled amounts listed inFormulations A and B represent a range that incorporates manufacturingvariations in content uniformity, manufacturing overages and loss ofpotency over the shelf-life period that are within the prescribedregulatory limits established by regulatory bodies such as the UnitedStates Food and Drug Administration and the United States Pharmacopeia.

All patents, patent applications, provisional applications, andpublications referred to or cited herein are incorporated by referencein their entirety, including all figures and tables, to the extent theyare not inconsistent with the explicit teachings of this specification.

Other embodiments are set forth within the following claims.

1.-37. (canceled)
 38. A method for reducing HbA1c levels in a mammalianpatient in need thereof, the method comprising: administering to thepatient a therapeutically effective amount of a composition comprising athiol-containing alkyl fatty acid formulation configured to reduce HbA1clevels in mammalian subjects, the thiol-containing alkyl fatty acidformulation comprising: alpha-lipoic acid, L-carnitine, CoQ10,L-glutamine, and L-arginine.
 39. The method of claim 38, wherein thethiol-containing alkyl fatty acid formulation comprises 600 mgalpha-lipoic acid, 400 mg L-carnitine, 100 mg CoQ10, 750 mg L-glutamine,and 1500 mg L-arginine.
 40. The method of claim 38, wherein thealpha-lipoic acid is present as a mixture of R and S isomers.
 41. Themethod of claim 38, wherein the mammalian patient is suffering from ametabolic dysfunction, and wherein the metabolic dysfunction ismetabolic syndrome, pre-diabetes, or diabetes.
 42. The method of claim38, wherein the composition comprising the thiol-containing alkyl fattyacid formulation is administered orally or intravenously.
 43. The methodof claim 38, wherein the mammalian patient, prior to administration ofthe composition, has an HbA1c level of 5.7% or greater.
 44. The methodof claim 38, wherein the alpha-lipoic acid, L-carnitine, CoQ10,L-glutamine, and L-arginine are active ingredients.
 45. The method ofclaim 44, wherein the composition further comprises pharmaceuticallyacceptable carriers and excipients.
 46. The method of claim 38, whereinthe composition is administered daily for one week or more.
 47. Themethod of claim 38, wherein the composition is administered one, two,three, four, or five times per day.
 48. The method of claim 38, whereinthe composition is administered weekly, bi-weekly, tri-weekly, ormonthly.
 49. The method of claim 38, wherein the mammalian patient ishuman.
 50. A thiol-containing alkyl fatty acid formulation for reducingHbA1c levels in mammalian subjects in need thereof comprisingalpha-lipoic acid, L-carnitine, CoQ10, L-glutamine, and L-arginine asactive ingredients and at least one pharmaceutically acceptable carrieror excipient.
 51. The thiol-containing alkyl fatty acid formulation ofclaim 50, further comprising a buffer.
 52. The thiol-containing alkylfatty acid formulation of claim 51, wherein the buffer minimizes rapiddegradation, increases absorption of the formulation, and/or minimizesstomach irritation or acid reflux.
 53. The thiol-containing alkyl fattyacid formulation of claim 51, wherein the buffer including ion-pairingsalts.
 54. The thiol-containing alkyl fatty acid formulation of claim50, wherein: the alpha-lipoic acid is present from about 5 wt % to about20 wt % of the formulation; the L-carnitine is present from about 5 wt %to about 20 wt % of the formulation; the CoQ10 is present from about 1wt % to about 5 wt % of the formulation; the L-glutamine is present fromabout 20 wt % to about 30 wt % of the formulation; and the L-arginine ispresent from at least about 31 wt % to about 65 wt % of the formulation.55. The thiol-containing alkyl fatty acid formulation of claim 50,wherein: the alpha-lipoic acid is present from about 600 mg to about 800mg; the L-carnitine is present from about 400 mg to about 450 mg; theCoQ10 is present from about 100 mg to about 150 mg; the L-glutamine ispresent from about 600 mg to about 800 mg; and the L-arginine is presentfrom about 1400 mg to about 1600 mg.
 56. The thiol-containing alkylfatty acid formulation of claim 50, wherein: a ratio of the alpha-lipoicacid to the L-carnitine in the formulation is between about 2.1:1 andabout 1.1:1; a ratio of the alpha-lipoic acid to the CoQ10 in theformulation is between about 8:1 and about 3:1; a ratio of thealpha-lipoic acid to the L-glutamine in the formulation is between about1:1 to about 1:3; and a ratio of the alpha-lipoic acid to the L-argininein the formulation is between about 1:1 to about 1:3.
 57. A method ofreducing HbA1c levels in a patient comprising administering atherapeutically effective amount of the thiol-containing alkyl fattyacid formulation of claim 50.